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Method for analyzing protein-protein interaction at a single molecule level in a cellular environment and method for measuring activated protein concentration in a cell solution

机译:在细胞环境中以单分子水平分析蛋白质-蛋白质相互作用的方法和测量细胞溶液中活化蛋白质浓度的方法

摘要

A method of analyzing protein-protein interactions at a single molecular level is disclosed. The method of analyzing the interactions between first proteins and second proteins at the single molecular level includes: preparing at least two substrates, in which first protein-binding molecules that are biomolecules to be bound to the first proteins are attached to each of the substrates; inducing binding between the first proteins and the first protein-binding molecules on the first substrate and the second substrate, respectively, by supplying the first proteins included in the control group-cell to the first substrate among the two substrates and supplying the first proteins included in the experimental group-cell to the second substrate among the two substrates; supplying cell lysates of cells including the marker-tagged second proteins to the first substrate and the second substrate, respectively, when the first proteins and the first protein-binding molecules are bound to the first substrate and the second substrate, respectively; and comparing and analyzing the interactions between the first proteins and the second proteins on the first substrate and the second substrate in the supply of the cell lysates to the first substrate and the second substrate, respectively. For observing the interactions between the first proteins and the second proteins, the state of each cell and activation levels of the first proteins can be compared and analyzed by comparing after varying of a type of cells supplying the first proteins.
机译:公开了一种在单个分子水平上分析蛋白质-蛋白质相互作用的方法。在单个分子水平上分析第一蛋白质和第二蛋白质之间的相互作用的方法包括:制备至少两个底物,其中作为要结合至第一蛋白质的生物分子的第一蛋白质结合分子附着于每个底物;通过向两个底物中的第一底物供应对照组细胞中包括的第一蛋白质并供应两个底物中的第一蛋白质,分别诱导第一底物和第二底物上的第一蛋白质和第一蛋白质结合分子之间的结合。在实验组电池中,至两个基板中的第二基板;当第一蛋白质和第一蛋白质结合分子分别结合到第一底物和第二底物上时,将包括带有标记标签的第二蛋白质的细胞的细胞裂解物分别提供给第一底物和第二底物。比较和分析分别向第一底物和第二底物提供细胞裂解物时第一底物和第二底物上的第一蛋白质和第二蛋白质之间的相互作用。为了观察第一蛋白质和第二蛋白质之间的相互作用,可以通过在改变提供第一蛋白质的细胞的类型之后进行比较来比较和分析每个细胞的状态和第一蛋白质的活化水平。

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